System for conducting stained antigen tests



March 19, 1940. R, w. TERRY 2,194,131

vSYSTEM FOR CONDUCTING STAINED ANTIGEN TESTS Filed July 3o, 193e Patented Mar. A19, '1940 j recordation of blood 'and'.serurn' vtestsin biological` SYSTMFR i @ame-ad for Lfacilitating the making; "interpretation and determinations. In its more specific aspects, lthe present invention provides an irnprovedniethod foi-conducting and recording stainedfantifg'en` mias been @stom-ary mempioy 'white @pai plates or mirrors. "On vthese plates orinirrorsytlieblood of Yeach fo'w-l'tested is'fseparatelyadniixd with .the'stained antigen and thereactionsfvisuaUy "f notedtoy dtermine the presenceofagglutinationt Y traces offthe tests lpreviously"madel thereomso With-tlie usev of such vitreous testing 'platesfolr minera-washing thereof vis necessary afterea li operation to ,permit of their reuse. The cleaning orwa'shin'g of the plates completely iemovesfall that in'sofar'as the actual'evidence ofthe original test is'gcon'cerned, the-samebe'comeslostor der',

stroyed. As a result of this condition, thefp ultry' raiser is without actual proof or'evidence fthe resultsof theftests made and in practice ifsiorjced ylargely-to rely upon oral statements..` u

- fIn accordance with the present invention" antigen test'sare made directlyjon appropriate 'she'etsotpapen or-ther corres'pondirig'relatively` inexpensive materia-1,' sothat said sheetr if ing the makingof thetests thereon, may'beled* inv appropriate"receptacles and kept las a 'durable' permanent' record, and one capable 'nfl-mingl profv duced=1at any future timefto disclose hefactual results of the tests setforth"threon.""

For a further understanding of the invention, Y

reference is to be had to the following description and the accompanying drawing, wherein:

' The figure is a plan view of a testing and recording sheet for conducting biological testsy and made in accordance with the present invention.

As shown in the accompanying drawing, the test sheetk I, made of paper or other equivalent material, such as certain types of cardboards, tracing cloths, or synthetic lms, is markedy or printed to provide a plurality of substantially square areas, as indicated at 2. 1 have found that a good grade of paper, known `to the art as 'hledger sheet, is well suited for the purpose and,

in addition, is quite economical. The square areasv 2 may be consecutively numbered and provided v v with such other data as may be desired in determining the subjects of the tests. Blanksgare thea yTerry, Columbus-.Ohio "'Abplicatit J'iny 136, 1936i sei-iai .'Noiias,

' 10mm! (cintas-sn v lett v*the* .bottom 'otfa 'sheet for the vrecordation of ,other ,data ,pertinent toI the tests made' and the certificate o fthose ,conducting., the tests;y y;

In use, the paper sheet is preferablylplaced 'on va mildly-heatedsurface:andthe blood from .each

fowLadmixed'with stained antigen in'situVis deovera considerable area of-jeach` space so Vtreated so` .that vrthe results ofsuch'V ,tests-may; be readily determined by. yisual.-inspeClJion. Careshould be used in the selection ofthepaper upon which process Aof .inanufacture,A seem unsuitable, since apparently the acidsor bleaching liquids detrif mentally-'attract vthe antigeni and' record perma,l

nency. ..A 'Otherpapers .that v absorb the antigen too rapidly'. are 'not",suitable, nor. a re papers that containcertain earths,- as sizingy materials, since the latter seem;4 tolabsorbthegcolor from the antigen and Ileave darli` colored spots. Ledger rsheet paper, however, appears to. be entirely.v `satisfac-1 to ry`, 'although rna ny other.A rades may befsuitable.,`Thi s paper shows amin'imumotstain from gen .wheiefdrops. of antigen are left in contact f o'rfafew `n iinutes from acidity,l or at, ,least there i not enough acid left in 'the` paperto influence thevtest, .Intact, theref appearsl-.to'be no interference inqany way when'this Vparticular,grande of paper isused.. Thus' one mayf'mixaloop-full of blood -with antigen and sjpreadfit out in af circlefofyjapproximatelyone.. inch in, diameter on thespaces fof -.thepap el,;pro vide d"fo' r that purpose'. r .The .papersheet: may

'then .b'eauiie tigorusly lacked and-there would4 be little orno tendency on-the part of, the ma {Itjrispractically free terials to spread beyondtheir,originally'allotted positions. This is 'not Atrue with the use of the opal plates or mirrors, since if the latter are vigorously rocked, the separate tests appearing thereon readily tend to run together.

Those tests on which there are definite reactions show a rapid formation of flakes and the reaction is quite clear on the Apaper sheet. After armement, the paper begins to curl from being wet and a depression aswell as a few higher spots formed in each wet circle. By rocking the sheet, the larger flakes are caught `on the fibers of the paper and stay up on the higher portions. The

liquid drains away from ythese flakes into the depression and leaves the flakes quite prominent. Since the formationof these ne confusing flakes occurs a little later, they take place after the depression is made inthe paper and form down depression which does not often occur since because of their lightness they stay in suspension in the liquid longer, are not caught on the fibers of the paper and flow back in the depression. In this depression where the liquid is deep, these fine flakes are invisible. In other words, this procedure eliminates from view the fine confusing flakes and makes more prominent the reaction denoting flakes.

An important feature of the present invention 4 resides in the fact that when tests have been conducted on the sheets and the results thereof noted, the sheets are permitted to dry and may then be filed away, thus furnishing the hatcherymen, poultry raiser or regulatory oiiicials with a permanent indisputable record that a particular ock was tested and the results thereof. At the present time, whenthese tests are conducted on white plates or mirrors, they are frequently washed, thereby destroying all record of the tests. i

In the present invention; permanent records may be obtained without additional effort as the test films must be made whether they are washed away or retained. The value of this permanent record possesses great possibilities. For instance, it gives public authorities and others a method of checking as to whether or not a certain ock was tested yand the accuracy of the test without actually being present at the time the test was conducted; lit furnishes hatcherymen or veterinarians protection to settle disputes and oii'ers a method of comparing at leisure the correlation between whole blood and tube tests. These test papers are also entirely serviceable for use with rapid serum tests. These test papers may be advantageously used in connection with the abortion control work in cattle as well as pullorum diseases in fowls, or in the testing of human blood as in typing for transfusions.

If the antigen ismixed with fowls blood or blood serum which do not contain antibodies, no reaction takes place, the mixture remaining uniform as when rst mixed. On the other hand, if the antigen is mixed withfowls blood or blood serum containing antibodies against this germ, the antibodies will react on the dead germs of the antigen, causing them to` form in stained colored masses or clumps.- This clumping of the germs in the antigen is known as agglutination.

It is a fundamental physiological responsefor an animal to attempt to overcome an infection. One of the ways this is accomplished is by the development within the animals body of substances that are capable of combating the infecting agent. These combatve substances are called antibodies, and are present in various tissues and blood of the animal. Y

Then conducting tests for pullorum disease in fowls, it is these antibodies that are tested for and not the germs, since the germs themselves are not circulating in the blood. The material used to determine whether or not the fowls blood contains these antibodies is called antigen, and consists of a vast number of dead pullorum disease germs mixed in salt water and containing a preservative and a violet dye. Blood is withdrawn from a fowl to be tested and admixed with antigen in situ, the mixture being made on the squares provided on the test sheet I.

The operator, by noting whether or not agglutination is present, which may be easily determined through visual inspection, can quickly and accurately note an infection in the fowls tested. The infected fowls may be readily determined and removed, thus enabling the non-infected fowls comprising thevv balance of the flock to be maintained in a healthy condition.

By conducting these tests on the paper sheet providedr lby the present invention, a complete lasting record of each test is available. It may be produced at any future time to show the results ofvany given test or tests and is` thus of considerable value to the poultry raiser. The tests conducted on the sheets are quite decisively negative and` positive, and the slight undulation of the paper when Wet renders reaction floccules very distinct and most all of any fine confusing specks vof foreign particles invisible. Following the drying of the test papers, reactions recorded thereon are as easily observable as when wet or immediately after original application to the paper.l

After the tests have dried, it may be helpful in certain instances in reading the tests to place the papers before a strong light, which will develop the colors and show them in the same manner as when wet.

What is claimed is:

The method of conducting stained antigen tests for pullorum disease in Vfowls, which comprises producing a liquid mixture of stained antigen and the fowls blood to be tested on a localized area'of a sheet 0f Paper which, when wetted withsaid mixture in an amount for. test purposes, undulates or wrinkles and thereby retains agglutinated reactants on the higher regions of the undulated portions of the paper, and causing the mixture to dry on the paper to produce a lasting record thereon of said test.

ROBERT W. TERRY. 

